基于DNA条形码技术构建王朗国家级自然保护区陆生脊椎动物遗传资源数据库及物种鉴定  

The establishment of terrestrial vertebrate genetic resource bank and species identification based on DNA barcoding in Wanglang National Nature Reserve

作　　者：邢超 林依 周智强 赵联军[4] 蒋仕伟 林蓁蓁 徐基良[1] 詹祥江[2] Chao Xing;Yi Lin;Zhiqiang Zhou;Lianjun Zhao;Shiwei Jiang;Zhenzhen Lin;Jiliang Xu;Xiangjiang Zhan(College of Ecology and Nature Conservation,Beijing Forestry University,Beijing 100083;Key Laboratory of Animal Ecology and Conservation Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing 100101;University of Chinese Academy of Sciences,Beijing 100049;Wanglang National Nature Reserve Administration,Pingwu,Sichuan 622550;Pingwu Forestry and Grassland Bureau,Pingwu,Sichuan 622550)

机构地区：[1]北京林业大学生态与自然保护学院,北京100083 [2]中国科学院动物研究所动物生态与保护生物学重点实验室,北京100101 [3]中国科学院大学,北京100049 [4]四川王朗国家级自然保护区管理局,四川平武622550 [5]平武县林业和草原局,四川平武622550

出　　处：《生物多样性》2023年第7期83-95,共13页Biodiversity Science

基　　金：四川王朗国家级自然保护区物种库项目。

摘　　要：DNA条形码(DNA barcode)是基因组中较短的、种内变异相对稳定的基因序列,已经成为生物多样性保护研究中物种鉴定、生物多样性评估的有力手段之一。四川王朗国家级自然保护区地处青藏高原东缘,属于世界生物多样性热点地区,具有丰富的生物资源,在我国珍稀动物保护领域具有重要地位。目前保护区已累积了大量的陆生脊椎动物监测数据,但缺乏遗传资源本底调查和基础的遗传资源数据库。本研究基于DNA条形码技术,以四川王朗国家级自然保护区为主要研究区域,基于样线法和博物馆标本调研,对所采集的314份样品进行DNA条形码分析,共鉴定兽类、鸟类、两栖类18目35科74种,首次获得了王朗齿突蟾(Scutigerwanglangensis)的线粒体基因(COI、12S-16S、16S、Cytb)及核基因(RAG1)的条形码序列信息,并通过比较不同监测方法说明了DNA条形码技术在动物多样性调查中的应用前景。本研究基于DNA条形码技术最终获得了216份DNA条形码数据,初步建立了保护区陆生脊椎动物遗传资源数据库,该数据库将为评估保护区生物多样性提供基础信息,为动物保护和管理工作提供技术支持。Aims:DNA barcoding using the short,highly conserved regions of the genome to make species-level identifications,is widely used in the species identification and diversity assessment of plants and animals in biodiversity conservation research.Wanglang National Nature Reserve is a global biodiversity hot spot located in the eastern part of Qinghai-Tibet Plateau with high level of biodiversity and has a leading role in the wildlife conservation in China.Although a lot of survey data of species diversity have been collected,genetic resource of terrestrial vertebrates in the reserve remains largely unclear.To address this,our study aims to establish a DNA barcoding database for terrestrial vertebrate species for the reserve.Methods:We collected genetic samples mainly using non-invasive sampling along the line transects in Wanglang National Nature Reserve and from museum specimens.We extracted DNA from the samples to obtain the barcode sequences by PCR.Each DNA barcode sequences were blasted against NCBI database for species identification.For those sequences that we could not directly get the matches in the reference database,we constructed phylogenetic trees using MEGA 11.0 with the aim to identify the species based on the sequences of its relative species from the public database.Results:The total of 314 samples including tissues from carcasses,feathers,feces and eggshells were collected and 74 species were identified using 16 pairs DNA barcoding primers.The sequences of mitochondrial COI,Cytb,12S-16S,16S genes and sequences of nuclear RAG1 gene of Scutiger wanglangensis were firstly reported.Compared with other wildlife monitoring techniques such as camera traps,drift fences and pitfall traps in the same time interval,DNA barcoding is more efficient in all taxon groups and has better performance in bird monitoring.Finally,A terrestrial vertebrate genetic resource bank contains 314 substantial samples and 216 DNA barcodes has been established.Conclusion:Our study established an elementary DNA barcoding database of 7

关 键 词：DNA条形码 王朗国家级自然保护区 陆生脊椎动物 生物多样性 条形码数据库 

分 类 号：Q953[生物学—动物学]